Validation of a UV Spectrophotometric Method for Determination of Proteolytic Activity of Enzymatic Detergents

Enzymatic detergents are widely used in health services. These products should be controlled to ensure their activity. In this paper, a UV spectrophotometric method for the determination of proteolytic activity in enzymatic detergents was validated. An artificial substrate (azocasein) was used to quantify the enzyme activity at 340 nm. The results showed that the method was satisfactory in respect to its specificity, linearity, precision, accuracy and robustness. This method fulfills both ICH’s and Brazilian criteria, demonstrating its suitability for routine analysis as well as its potential application for regulatory purposes.     

The Role of F-Box Proteins during Viral Infection

The F-box domain is a protein structural motif of about 50 amino acids that mediates protein–protein interactions. The F-box protein is one of the four components of the SCF (SKp1, Cullin, F-box protein) complex, which mediates ubiquitination of proteins targeted for degradation by the proteasome, playing an essential role in many cellular processes. Several discoveries have been made on the use of the ubiquitin–proteasome system by viruses of several families to complete their infection cycle. On the other hand, F-box proteins can be used in the defense response by the host. This review describes the role of F-box proteins and the use of the ubiquitin–proteasome system in virus–host interactions.     

Experimental behaviour of the reverse channel joint component at elevated and ambient temperatures

The reverse channel connection appears to have the best combination of desirable features under fire loading: moderate construction cost, ability to develop catenary action and extremely high ductility through deformation of the web channel (Ding and Wang, 2007). This paper presents the results of an experimental investigation of a reverse channel component conducted at the University of Coimbra as part of the European RFCS COMPFIRE Project, the main focus of which is to characterise the behaviour of steel joints that connect steel beams to concrete-filled tubular columns under natural fire loading. A series of tensile and compressive tests at ambient and elevated temperatures was conducted. The purposes of the experimental tests were to characterise the strength, stiffness and ductility of this joint component and to establish a relationship between force, displacement and temperature.     

Characterization of peptide and amino acid profile in casein hydrolysates

The nutritional quality of protein hydrolysates has been related in several reports to their di- and tripeptide contents. In the present work different hydrolytic conditions were tested using papain in order to prepare casein hydrolysates with a suitable peptide profile for being used in special diets. The hydrolysates were fractionated by size-exclusion HPLC and the rapid Correct Fraction Area method was used for quantifying the peptides. Among the five hydrolytic conditions studied, three of them gave rise to preparations having nutritionally similar peptide profiles. However, the use of the temperature of 37 degrees C and enzyme:substrate ratio (E:S) of 2% may probably be the most economical condition for industrial production.     

Structural analysis of Si-substituted hydroxyapatite: zeta potential and X-ray photoelectron spectroscopy

The aim of this study was to determine the effect of the incorporation of silicon on the surface charge of hydroxyapatite (HA) and to assess surface structural changes of HA and Si–HA induced by dissolution in both static and dynamic systems. X-ray photoelectron spectroscopy (XPS) analysis showed that SiO₄ ^4– groups were substituted for PO₄ ^3– groups in the silicon-hydroxyapatite (Si–HA) lattice according to a previously proposed substitution mechanism without the formation of other crystalline phases, such as tricalcium phosphate or calcium oxide. The substituted silicon induced a decrease in the net surface charge and the isoelectric point of HA as determined by zeta potential (ZP) measurements. At physiological pH=7.4 the surface charge of Si–HA was significantly lowered compared to unmodified HA, i.e. –50±5 to –71±5 eV, caused by the presence of silicate groups in the HA lattice, which may account for a faster in vitro apatite formation using SBF testing. XPS results indicated that silicon seems to be preferentially leached out from Si–HA surface compared to other ionic species after dissolution studies in tris-buffer using a dynamic system.     

Developmental stability in stocks of White Leghorn chickens

The degree of asymmetry in bilateral morphological characters may reflect genetic and environmental stressors. Shank length and diameter, weight and length of the first primary wing feather, and distance between the junction of upper and lower mandibles and auditory canal (face length) were used to classify bilateral types and measure relative asymmetry (RA) in six genetic stocks. The stocks were the S23 generation of White Leghorn lines selected for high or low antibody response to SRBC, sublines in which selection had been relaxed for eight generations, and reciprocal crosses of the selected lines. Differences were found among all stocks for the traits measured. Rankings among traits for RA in descending order were face length, shank diameter, feather weight, and shank and feather lengths. The RA of shank and feather lengths did not differ from each other. An overall RA composed of mean RA of the five traits showed that the two selected lines exhibited greater RA than the crosses between them. The RA of the two lines where selection had been relaxed was similar to that of selected lines. This research suggests that an overall RA created as a combination of RA of several bilateral traits can be a valid measure of genetic stress in chickens and provides a method of comparing developmental stability among populations.     

The t(5;17) variant of acute promyelocytic leukemia expresses a nucleophosmin-retinoic acid receptor fusion

The classical action of the hormone 1,25-dihydroxyvitamin D3 (VD) is the regulation of calcium metabolism. In contrast, the peptide hormone atrial natriuretic factor (ANF) is one of the few known nonclassical VD responding genes. We screened the promoter of the rat ANF gene and identified a typical VD receptor (VDR) binding site formed by a direct repeat of two hexameric core binding motifs spaced by three nucleotides, between positions -907 and -891. Like most of the DR3-type VD response elements this sequence is bound with high affinity (Kd = 0.53 nM) by a heterodimer formed by VDR and retinoid X receptor. In a heterologous promoter context one copy of this sequence mediated an about fourfold gene activation by VD and a half-maximal activation (EC50) value of 0.48 nM VD. This characterizes the identified sequence as one of the most potent VD response elements.     

XR-C1, a new CHO cell mutant which is defective in DNA-PKcs, is impaired in both V(D)J coding and signal joint formation

DNA-dependent protein kinase (DNA-PK) plays an important role in DNA double-strand break (DSB) repair and V(D)J recombination. We have isolated a new X-ray-sensitive CHO cell line, XR-C1, which is impaired in DSB repair and which was assigned to complementation group 7, the group that is defective in the XRCC7 / SCID ( Prkdc ) gene encoding the catalytic subunit of DNA-PK (DNA-PKcs). Consistent with this complementation analysis, XR-C1 cells lackeddetectable DNA-PKcs protein, did not display DNA-PK catalytic activity and were complemented by the introduction of a single human chromosome 8 (providing the Prkdc gene). The impact of the XR-C1 mutation on V(D)J recombination was quite different from that found in most rodent cells defective in DNA-PKcs, which are preferentially blocked in coding joint formation, whereas XR-C1 cells were defective in forming both coding and signal joints. These results suggest that DNA-PKcs is required for both coding and signal joint formation during V(D)J recombination and that the XR-C1 mutant cell line may prove to be a useful tool in understanding this pathway.     

Influence of sphingolipids on T lymphocyte activation

Two experiments are reported which examined the viability of motor output hypothesis as an explanation for manual asymmetries in goal-directed movement. Experiment 1 isolated the variability due to force generation by directly assessing precision of force production during an isometric wrist flexion task. Experiment 2 examined the additional role of externally based and internally created timing patterns on the performance of a repetitive force production task. Virtually no effects involving hand were apparent in either experiment. These findings provide no support for a hypothesis based solely on motor output to adequately account for hand differences in the performance of rapid, goal-directed movement.